LOCUS       Exported                9748 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pYL15, complete sequence.
ACCESSION   KU378202
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9748)
  AUTHORS   Pomraning KR.
  TITLE     localization of lipid biogenesis in yarrowia lipolytica
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9748)
  AUTHORS   Pomraning KR.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JAN-2016) Environmental Molecular Science Lab, Pacific
            Northwest National Lab, 902 Battelle Blvd, Richland, WA 99352, USA
REFERENCE   3  (bases 1 to 9748)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9748
                     /organism="Cloning vector pYL15"
                     /mol_type="other DNA"
                     /db_xref="taxon:1890043"
     CDS             10..753
                     /codon_start=1
                     /product="10xGly-sGFP"
                     /label=10xGly-sGFP
                     /note="10x Glycine linker attached to super-fold green 
                     fluorescent protein"
                     /protein_id="APB62234.1"
                     /translation="GGGGGGGGGGRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGD
                     ATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQE
                     RTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITA
                     DKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNE
                     KRDHMVLLEFVTAAGITHGMDELYK"
     CDS             40..750
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     gene            complement(766..3029)
                     /gene="leu2"
                     /label=leu2
     CDS             complement(1028..2245)
                     /codon_start=1
                     /gene="leu2"
                     /product="LEU2"
                     /label=leu2
                     /protein_id="APB62232.1"
                     /translation="MEPETKKTKTDSKKIVLLGGDFCGPEVIAEAVKVLKSVAEASGTE
                     FVFEDRLIGGAAIEKEGEPITDATLDICRKADSIMLGAVGGAANTVWTTPDGRTDVRPE
                     QGLLKLRKDLNLYANLRPCQLLSPKLADLSPIRNVEGTDFIIVRELVGGIYFGERKEDD
                     GSGVASDTETYSVPEVERIARMAAFLALQHNPPLPVWSLDKANVLASSRLWRKTVTRVL
                     KDEFPQLELNHQLIDSAAMILIKQPSKMNGIIITTNMFGDIISDEASVIPGSLGLLPSA
                     SLASLPDTNEAFGLYEPCHGSAPDLGKQKVNPIATILSAAMMLKFSLNMKPAGDAVEAA
                     VKESVEAGITTADIGGSSSTSEVGDLLPTRSRSCSRRSKSFLRRIDGRSKLTRLRVGLP
                     AGSASV"
     misc_feature    3037..3281
                     /note="modified version of ARS68, contains centromere and 
                     origin of replication sequences for Yarrowia lipolytica"
     rep_origin      3456..4123
     rep_origin      complement(3512..4100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4274..5068)
                     /codon_start=1
                     /gene="kanR"
                     /product="kanR"
                     /label=kanR
                     /protein_id="APB62233.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(4274..5068)
                     /gene="kanR"
                     /label=kanR
     promoter        complement(5069..5173)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5489..5505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5516..5534
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5539..5550
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      8995..9739
                     /regulatory_class="promoter"
                     /gene="exp1"
     gene            8995..9739
                     /gene="exp1"
                     /label=exp1